Evaluation of pan-dermatophyte nested PCR in diagnosis of onychomycosis in comparison with direct microscopy and culture

This work aimed to compare nested PCR using novel primers targeting the pan-dermatophyte-specific sequence of the chitin synthase 1 gene [CHS1] with KOH microscopy and culture isolation for diagnosis of clinically suspected onychomycosis. This study was conducted during the period from December, 2012 to October 2013. Forty patients attending Outpatient Dermatology and Andrology Clinic in Benha University Hospital. This study was done on forty patients 15 cases were female and the other 25 cases were males with abnormal nails. Their ages ranged from 22 to 77 years. As many as 19 patients were living in rural areas, while 21 patients came from urban areas. Nail scrapings were collected and examined using direct KOH microscopic examination, culture and PCR using double sets of primers. As regard direct microscopy by KOH examination; 33 [82.50%] cases were positive, while 7 [17.5%] were negative. Culture was positive only in 19[47.5%] of nail samples revealing different fungi. Dermatophytes were isolated from 15[37.5%] cases; most of them were T. mentagrophytes. And in 4 cases the only isolated non dermatophytic organism was Aspergillus Niger spp. [10.00%]. Nested PCR was positive in 26 [65.00%] nail samples. It is concluded that nested PCR targeting the CHS1 gene may be considered the gold standard for detection of dermatophytes in patients with onychomycosis and can aid the clinician in initiating prompt and appropriate antifungal therapy. PCR is a very powerful tool for microbiology and clinical mycology. It can detect very small amounts of nucleic acids. This technique may also play an important role in large-scale studies and in the management of problematic cases of onychopathies

[Prevalence and antibiotic resistance pattern of Staphylococcus aureus strains isolated from food stuff]

Staphylococcus aureus is considered to be one of the leading causes of food-borne illnesses. Foodstuff contamination may occur directly from contaminated food-producing animals or may result from poor hygiene during food production processes, or the retail and storage of foods, since humans may carry the microorganism. The number of S. aureus strains that exhibits antimicrobial-resistance properties has increased, together with the potential risk of transmitting the same properties to the human micro flora via food or inducing infections hard to be treated. The aim of this study was to evaluate the occurrence of S. aureus in various food samples and determination of antibiotic resistance pattern in this isolates. A total of 1047 food samples were analysed from July 2006 to December 2007. To determine the presence of S.aureus, the samples were analysed according to the guidelines of Iran standard instructions [No. 1194]. S.aureus isolates were tested for susceptibility to a panel of 11 antimicrobics using the agar disc diffusion method on Muller-Hinton agar. Of 1047 samples analysed 100 [9.5%] were contaminated with S.aureus. Among these contaminated samples, 31% showed antimicrobial resistance properties to at least one of the antibiotic tested and 15 antibiotypes were determined. According to the observed prevalence of S.aureus strains in food samples and their antibiotic resistance pattern, more attention should be paid in foodstuff industry to prevent contamination and transmission of resistant strains

[ survey on the presence of anti-GAD in type 1 diabetic patients and their first-degree relatives in comparison with healthy individuals]

Glutamic acid decarboxylase [GAD] catalyzes the conversion of glutamic acid to gamma-aminobutyric acid [GABA]. GAD65 isozyme is present in the pancreatic beta-cells. In the prediabetes period and during the beta-cell destruction, GAD is released as an autoantigen and anti-GAD autoantibodies appear in serum. Islet Cell Autoantibodies[ICAs] including anti-GAD are detectable in serum of diabetic patients up to 10 years before appearance of diabetes symptoms. This is an important predictive marker for diagnosis of prediabetic patients, especially in the first-degree relatives of diabetic patients for genetic factors. Anti-GAD is an important marker for detection of beta-cells destruction. The patients with high titers of anti-GAD have a worse disease prognosis and are in greater need of insulin injections. This survey is a case-control study aimed at detection of anti-GAD presence in sera of type 1 diabetic patients and their first-degree relatives and comparison with healthy individuals. Fifty type 1 diabetic patients with mean age of 12.24 +/- 6.2 years and mean disease duration of 34.5 +/- 8.4 months, 35 first-degree relatives and 50 normal individuals without familial diabetes were included in the study; all the individuals were chosen by a random sampling method. The values of fasting blood sugar were determined in first-degree relatives and controls and all were found to be normal. The values of anti-GAD were determined by ELISA method. Median values of anti-GAD in cases and controls were 28, [range: 5-2700] ng/ml and 2, [0-10] ng/ml, respectively. The anti-GAD titers were significantly higher in patients than in normal individuals and relatives together [p<0.0001]. Median value of anti-GAD in first-degree relatives was 7, [0-950] ng/ml. There was a significant statistical difference between anti-GAD titers in first-degree relatives and controls, [p<0.01]. There was a significant difference between mean value of age and diabetes duration in anti-GAD positive and anti-GAD negative patients, [p<0.05]. There was a negative correlation between anti-GAD and age, diabetes duration, disease beginning age of patients, [r=-0.155,-0.158,-0.036], respectively. By increasing of anti-GAD in diabetic patients and their first-degree relatives it may be concluded that measurement of anti-GAD is an important and beneficial tool for detection and diagnosis of prediabetic and diabetic patients

Study of some pathophysiological factors in children with acquired aplastic anaemia

Aplastic anaemia [AA] is a heterogenous disease including different pathophysiological conditions, characterized by severely diminished numbers of bone marrow [B.M] haematopiotic cells resulting in failure of the marrow to produce mature blood elements, this study aimed to determine the role of apoptosis, tumour necrosis factor alpha [TNFalpha.] and granulocytes-macrophage colony stimulating factor [GM-CSF] in the pathogensis of AA. Thirty five cases diagnosed as acquired A.A, Twenty cases newly diagnosed, 8 cases in partial remission and 7 cases in complete remission were studied. Twenty age and sex matched children were taken as control group.Mononuclear cell apoptosis was estimated using flowcytometry by TUNEL method.Serum level of TNFa. and GM-CSF were estimated using ELISA technique. The result of this study revealed significant increase of mononuclear cell apoptosis and TNFalpha in newly diagnosed cases of AA and in cases with partial remission more than in control group. Also there was an insignificant increase of apoptosis in the cases in complete remission than in control group. Serum GM-CSF was significantly reduced in all cases of aplastic anaemia except cases in complete remission when compared with control group.There was inverse correlation between mononuclear cell apoptosis,TNFalpha, and GM-CSF [r - 0.52, - 0.55 respectively, p <0.001] In conclusion increase the rate of mononuclear cell apoptosis,TNF-alpha and decreased level of GM-CSF play a role in pathophysiology of B.M failure and their follow up may be one of the important parameters to assure complete recovery